Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum

Schleicher, Rosemary L.; Encisco, Sara E.; Chaudhary‐Webb, Madhulika; Paliakov, Ekaterina; McCoy, Leslie F.; Pfeiffer, Christine M.

doi:10.1016/j.cca.2011.05.010

Cited by 101 publications

(82 citation statements)

References 18 publications

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“…2 The inversion of stereochemistry at C3 is chemically distant from the triene chromophore, but differences in UV absorption cannot be excluded. However, we would not expect the difference in UV absorption to exceed the < 10% difference between…”

Section: Discussionmentioning

confidence: 99%

Enhanced 3-epi-25-hydroxyvitamin D₃ signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

2013

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Background: 3-epi-25-hydroxyvitamin D 3 (3-epi-25OHD 3 ) interferes in most liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays for 25-hydroxyvitamin D (25OHD). The clinical significance of this is unclear, with concentrations from undetectable to 230 nmol/L reported. Many studies have quantified 3-epi-25OHD 3 based on 25OHD 3 calibrators or other indirect methods, and we speculated that this contributes to the observed variability in reported 3-epi-25OHD 3 concentrations. Methods: We compared continuous MS/MS infusions of 3-epi-25OHD 3 and 25OHD 3 solutions, spiked both analytes into the same serum matrix and analysed patient samples to assess the effect of three different quantitation methods on 3-epi-25OHD 3 concentration. Experiments were performed on an LC-MS/MS system using a phenyl column which does not resolve 3-epi-25OHD 3 , and a modified method utilizing a Zorbax SB-CN column that chromatographically resolves 3-epi-25OHD 3 from 25OHD 3 . Results: A greater 3-epi-25OHD 3 signal, compared with 25OHD 3 , was observed during equimolar post-column continuous infusion of analyte solutions, and following analysis of a serum pool spiked with both analytes. 3-epi-25OHD 3 signal enhancement was dependent on mobile phase composition. Compared with 3-epi-25OHD 3 calibrators, indirect quantitation methods resulted in up to 10 times as many samples having 3-epi-25OHD 3 concentrations ! 10 nmol/L, and an approximately fourfold increase in the maximum observed 3-epi-25OHD 3 concentration to 95 nmol/L. Conclusions: Enhanced 3-epi-25OHD 3 signal leads to overestimation of its concentrations in the indirect quantitation methods used in many previous studies. The enhanced signal may contribute to greater interference in some 25OHD LC-MS/MS assays than others. We highlight that equimolar responses cannot be assumed in LC-MS/MS systems, even if two molecules are structurally similar.

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Section: Discussionmentioning

confidence: 99%

Enhanced 3-epi-25-hydroxyvitamin D₃ signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

2013

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“…Furthermore, additional complications arise from isomeric contributions, where the C-3 epimer of 25(OH)D 3 presents a particular challenge [2]. Whether or not the epimeric species are biologically relevant is currently unknown [7]; regardless, chromatographic separation is essential to avoid bias in the analysis [1,2,8,9].…”

Section: Introductionmentioning

confidence: 99%

Triple Quadrupole Versus High Resolution Quadrupole-Time-of-Flight Mass Spectrometry for Quantitative LC-MS/MS Analysis of 25-Hydroxyvitamin D in Human Serum

Geib

Sleno

Hall

et al. 2016

J. Am. Soc. Mass Spectrom.

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Abstract. We describe a systematic comparison of high and low resolution LC-MS/ MS assays for quantification of 25-hydroxyvitamin D 3 in human serum. Identical sample preparation, chromatography separations, electrospray ionization sources, precursor ion selection, and ion activation were used; the two assays differed only in the implemented final mass analyzer stage; viz. high resolution quadrupolequadrupole-time-of-flight (QqTOF) versus low resolution triple quadrupole instruments. The results were assessed against measured concentration levels from a routine clinical chemiluminescence immunoassay. Isobaric interferences prevented the simple use of TOF-MS spectra for extraction of accurate masses and necessitated the application of collision-induced dissociation on the QqTOF platform. The two mass spectrometry assays provided very similar analytical figures of merit, reflecting the lack of relevant isobaric interferences in the MS/MS domain, and were successfully applied to determine the levels of 25-hydroxyvitamin D for patients with chronic liver disease.

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“…The vitamin D metabolites generally elute in the order trihydroxylated < dihydroxylated < monohydroxylated metabolites, that is, for vitamin D 3 -related molecules, the order of chromatographic retention is 1,25(OH) 2 Possibly more interesting-from a chromatography point of view-are the biochemically formed isomers after stereochemical reversal (β→α) at C-3; for example, the epimers 25(OH)D 3 and 3-epi-25(OH)D 3 . The subtle diastereomeric differences between these species can be distinguished using specialized columns, such as combined C18/chiral [50], pentafluorophenyl (PFP) [51][52][53], and cyano (CN) [54][55][56][57].…”

Section: Liquid Chromatographic Separation Of Vitamin D Metabolitesmentioning

confidence: 99%

Synthesis of Low Abundant Vitamin D Metabolites and Assaying Their Distribution in Human Serum by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) as a New Tool for Diagnosis and Risk Prediction of Vitamin DRelated Diseases

Kattner¹,

Volmer²

2017

A Critical Evaluation of Vitamin D - Basic Overview

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This chapter provides an overview of versatile and efficient chemical syntheses of vitamin D derivatives by application of either linear or convergent synthesis approaches. Synthesis of the most relevant naturally occurring vitamin D metabolites and their deuterated counterparts to use as calibration and reference standards in LC-MS/MS assays is also shown. The chapter then summarizes the most important mass spectrometric approaches to quantify important vitamin D metabolites in human biofluids. In addition, new developments are described that are aimed at the pathobiological interpretation of the measured vitamin D metabolite distributions in various human diseases.

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Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum

Cited by 101 publications

References 18 publications

Enhanced 3-epi-25-hydroxyvitamin D₃ signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

Enhanced 3-epi-25-hydroxyvitamin D₃ signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

Triple Quadrupole Versus High Resolution Quadrupole-Time-of-Flight Mass Spectrometry for Quantitative LC-MS/MS Analysis of 25-Hydroxyvitamin D in Human Serum

Synthesis of Low Abundant Vitamin D Metabolites and Assaying Their Distribution in Human Serum by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) as a New Tool for Diagnosis and Risk Prediction of Vitamin DRelated Diseases

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Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum

Cited by 101 publications

References 18 publications

Enhanced 3-epi-25-hydroxyvitamin D3 signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

Enhanced 3-epi-25-hydroxyvitamin D3 signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

Triple Quadrupole Versus High Resolution Quadrupole-Time-of-Flight Mass Spectrometry for Quantitative LC-MS/MS Analysis of 25-Hydroxyvitamin D in Human Serum

Synthesis of Low Abundant Vitamin D Metabolites and Assaying Their Distribution in Human Serum by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) as a New Tool for Diagnosis and Risk Prediction of Vitamin DRelated Diseases

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Enhanced 3-epi-25-hydroxyvitamin D₃ signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays

Enhanced 3-epi-25-hydroxyvitamin D₃ signal leads to overestimation of its concentration and amplifies interference in 25-hydroxyvitamin D LC-MS/MS assays